scholarly journals Immunohistochemical localization of active caspase-3 in the mouse ovary: growth and atresia of small follicles

Reproduction ◽  
2002 ◽  
pp. 659-665 ◽  
Author(s):  
MA Fenwick ◽  
PR Hurst
Keyword(s):  
Granulosa Cells ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Active Form ◽  
Cysteine Proteases ◽  
Immunohistochemical Localization ◽  
Mouse Ovary ◽  
Physiological Factors ◽  
Cell Suicide ◽  
Active Caspase

Caspase-3 belongs to a family of highly conserved cysteine proteases that mediate the course of apoptotic cell suicide. It is recognized that ovarian follicular atresia is associated with apoptosis, a process that has been characterized mainly in larger antral follicles. The aims of this study were to investigate the expression of caspase-3 in the mouse ovary, and determine whether active caspase-3 is present within smaller follicles, which may constitute the resting pool. The inactive enzyme was expressed as a 32 kDa band on a western blot of tissue extracts, whereas the active form was localized immunohistochemically. Bromodeoxyuridine (BrdU) was administered to mice (n = 7) during a 12 h period and subsequently localized to identify potentially quiescent follicles. Measurements of BrdU-positive cells in the mouse ovary were extrapolated with data obtained by morphometric analyses of small follicles using the nucleator technique. BrdU was incorporated into the granulosa cells of follicles regardless of size and the number of cells they contained, but was absent in a large proportion (89%) of small, single layered follicles. Active caspase-3 was localized to both the oocyte and granulosa cells of follicles that were considered to be undergoing atresia, but was not localized to the granulosa cells of any small, single layered follicles. The results of this study indicate that, in small follicles, granulosa cell proliferation occurs independently of the size of follicles and the number of constituent cells, and that follicles of this type may be inherently less susceptible to the normal physiological factors that induce atresia.

Apoptosis and active caspase-3 expression in human granulosa cells

2005 ◽  
Vol 83 (2) ◽  
pp. 426-431 ◽  
Author(s):  
Violeta Glamočlija ◽  
Katarina Vilović ◽  
Mirna Saraga-Babić ◽  
Anamarija Baranović ◽  
Damir Sapunar
Keyword(s):  
Granulosa Cells ◽  
Caspase 3 ◽  
Human Granulosa Cells ◽  
Active Caspase

scholarly journals Interferon-Tau Exerts Direct Prosurvival and Antiapoptotic Actions in Luteinized Bovine Granulosa Cells

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Raghavendra Basavaraja ◽  
Senasige Thilina Madusanka ◽  
Jessica N. Drum ◽  
Ketan Shrestha ◽  
Svetlana Farberov ◽  
...  
Keyword(s):  
Cell Viability ◽  
Granulosa Cells ◽  
Early Pregnancy ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Interferon Tau ◽  
Cell Counts ◽  
Domestic Ruminants ◽  
Stat1 Protein

Abstract Interferon-tau (IFNT), serves as a signal to maintain the corpus luteum (CL) during early pregnancy in domestic ruminants. We investigated here whether IFNT directly affects the function of luteinized bovine granulosa cells (LGCs), a model for large-luteal cells. Recombinant ovine IFNT (roIFNT) induced the IFN-stimulated genes (ISGs; MX2, ISG15, and OAS1Y). IFNT induced a rapid and transient (15–45 min) phosphorylation of STAT1, while total STAT1 protein was higher only after 24 h. IFNT treatment elevated viable LGCs numbers and decreased dead/apoptotic cell counts. Consistent with these effects on cell viability, IFNT upregulated cell survival proteins (MCL1, BCL-xL, and XIAP) and reduced the levels of gamma-H2AX, cleaved caspase-3, and thrombospondin-2 (THBS2) implicated in apoptosis. Notably, IFNT reversed the actions of THBS1 on cell viability, XIAP, and cleaved caspase-3. Furthermore, roIFNT stimulated proangiogenic genes, including FGF2, PDGFB, and PDGFAR. Corroborating the in vitro observations, CL collected from day 18 pregnant cows comprised higher ISGs together with elevated FGF2, PDGFB, and XIAP, compared with CL derived from day 18 cyclic cows. This study reveals that IFNT activates diverse pathways in LGCs, promoting survival and blood vessel stabilization while suppressing cell death signals. These mechanisms might contribute to CL maintenance during early pregnancy.

scholarly journals Exoenzyme Y induces extracellular active caspase-7 accumulation independent from apoptosis: modulation of transmissible cytotoxicity

2020 ◽  
Vol 319 (2) ◽  
pp. L380-L390
Author(s):  
Phoibe Renema ◽  
Natalya Kozhukhar ◽  
Viktoriya Pastukh ◽  
Domenico Spadafora ◽  
Sunita Subedi Paudel ◽  
...  
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Cell Swelling ◽  
Wild Type ◽  
Cell Infection ◽  
Heat Stable ◽  
Caspase 7 ◽  
Executioner Caspases ◽  
Active Caspase

Caspase-3 and -7 are executioner caspases whose enzymatic activity is necessary to complete apoptotic cell death. Here, we questioned whether endothelial cell infection leads to caspase-3/7-mediated cell death. Pulmonary microvascular endothelial cells (PMVECs) were infected with Pseudomonas aeruginosa (PA103). PA103 caused cell swelling with a granular appearance, paralleled by intracellular caspase-3/7 activation and cell death. In contrast, PMVEC infection with ExoY+ (PA103 Δ exoUexoT::Tc pUCP exoY) caused cell rounding, but it did not activate intracellular caspase-3/7 and it did not cause cell death. However, ExoY+ led to a time-dependent accumulation of active caspase-7, but not caspase-3, in the supernatant, independent of apoptosis. To study the function of extracellular caspase-7, caspase-7- and caspase-3-deficient PMVECs were generated using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology. Caspase-7 activity was significantly reduced in supernatants from infected caspase-7-deficient cells but was unchanged in supernatants from infected caspase-3 deficient cells, indicating an uncoupling in the mechanism of activation of these two enzymes. Because ExoY+ leads to the release of heat stable amyloid cytotoxins that are responsible for transmissible cytotoxicity, we next questioned whether caspase-7 contributes to the severity of this process. Supernatants obtained from infected caspase-7-deficient cells displayed significantly reduced transmissible cytotoxicity when compared with supernatants from infected wild-type controls, illustrating an essential role for caspase-7 in promoting the potency of transmissible cytotoxicity. Thus, we report a mechanism whereby ExoY+ infection induces active caspase-7 accumulation in the extracellular space, independent of both caspase-3 and cell death, where it modulates ExoY+-induced transmissible cytotoxicity.

scholarly journals Active caspase 3 and DNA fragmentation as markers for apoptotic cell death in primary and metastatic liver tumours

Pathology ◽  
2007 ◽  
Vol 39 (6) ◽  
pp. 558-564 ◽  
Author(s):  
Eva Karamitopoulou ◽  
Luca Cioccari ◽  
Sabine Jakob ◽  
Claudio Vallan ◽  
Thomas Schaffner ◽  
...  
Keyword(s):  
Cell Death ◽  
Dna Fragmentation ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Apoptotic Cell Death ◽  
Liver Tumours ◽  
Metastatic Liver ◽  
Active Caspase

335. X-LINKED INHIBITOR OF APOPTOSIS PROTEIN (XIAP) EXPRESSION PATTERNS IN THE SHEEP OVARY

2010 ◽  
Vol 22 (9) ◽  
pp. 135 ◽  
Author(s):  
H. R. Douglas ◽  
I. C. Kokay ◽  
D. R. Grattan ◽  
P. R. Hurst
Keyword(s):  
Inverse Relationship ◽  
Caspase 3 ◽  
Expression Patterns ◽  
Cysteine Proteases ◽  
Follicular Atresia ◽  
Study Objective ◽  
Antral Follicles ◽  
Xiap Expression ◽  
Apoptosis Protein ◽  
Active Caspase

At reproductive age, the ovary undergoes continual cyclicity of follicles due to multiple positive and negative signals that promote follicle growth and development, selection for ovulation, or atresia. The majority of follicles undergo atresia, a degenerative process involving granulosa cell apoptosis. This process is executed by caspases, which are cysteine proteases. Caspases are potently inhibited by XIAP providing a potential mechanism to control follicular atresia. The study objective was to test the hypothesis that XIAP will show elevated expression in healthy antral follicles compared to atretic antral follicles and will show an inverse relationship with active caspase-3 immunoreactivity in the same antral follicle during the sheep estrous cycle. Reproductively mature Romney ewes (n = 9) had estrous cycles synchronized with a prostaglandin F2α analogue, then tissue was collected on days 14, 15 and 16 of the subsequent natural cycle. Analysis involved determining the presence and localization of XIAP using in situ hybridization histochemistry and immunohistochemistry then comparing XIAP expression patterns with distribution of active caspase-3 protein. XIAP mRNA was not detected in primordial, primary and secondary follicles. In contrast, XIAP protein was present from the primary stage onwards. Antral follicles showed positive XIAP mRNA and protein expression in both granulosa and thecal cell layers and antral follicles on the same tissue section showed variable expression. All day 14 antral follicles were positive for XIAP mRNA expression irrespective of the level of active caspase-3 immunoreactivity, whereas an inverse relationship between active caspase-3 and XIAP was apparent in the majority of day 15 and 16 antral follicles. XIAP protein was widely expressed in active caspase-3 negative antral follicles and indicated a negative correlation with the onset of active caspase-3 expression in the majority of follicles. These results indicate that XIAP may regulate follicular atresia and act as an indicator of follicular health in the sheep ovary.

scholarly journals Melatonin Alleviates Hypoxia-Induced Apoptosis of Granulosa Cells by Reducing ROS and Activating MTNR1B–PKA–Caspase8/9 Pathway

Antioxidants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 184
Author(s):  
Jing-Li Tao ◽  
Xuan Zhang ◽  
Jia-Qi Zhou ◽  
Cheng-Yu Li ◽  
Ming-Hui Yang ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Heme Oxygenase 1 ◽  
Inhibitory Effects ◽  
Glutathione S Transferase ◽  
Competitive Antagonist ◽  
Hypoxia Exposure ◽  
Induced Apoptosis

In mammalian ovaries, the avascular environment within follicular cavity is supposed to cause hypoxic status in granulosa cells (GCs), leading to apoptotic cell death accompanied by cumulative reactive oxygen species (ROS) production. Melatonin (N-acetyl-5-methoxytryptamine, MT), a broad-spectrum antioxidant that exists in porcine follicle fluid, was suggested to maintain GCs survival under stress conditions. In this study, using the established hypoxic model (1% O2) of cultured porcine GCs, we explored the effect of MT on GCs apoptosis. The results showed that MT restored cell viability and reduced the apoptosis of GCs during hypoxia exposure. In addition, GCs treated with MT exhibited decreased ROS levels and increased expression of antioxidant enzymes including heme oxygenase-1 (HO-1), glutathione S-transferase (GST), superoxide dismutase 1 (SOD1), and catalase (CAT) upon hypoxia incubation. Moreover, the hypoxia-induced expression of cleaved caspase 3, 8, and 9 was significantly inhibited after MT treatment. In contrast, blocking melatonin receptor 2 (MTNR1B) with a competitive antagonist 4-phenyl-2-propionamidotetralin (4P-PDOT) diminished the inhibitory effects of MT on caspase 3 activation. By detecting levels of protein kinase (PKA), a downstream kinase of MTNR1B, we further confirmed the involvement of MT–MTNR1B signaling in mediating GCs protection during hypoxia stress. Together, the present data provide mechanistic evidence suggesting the role of MT in defending GCs from hypoxia-induced apoptosis.

scholarly journals Effects of Silver Nanoparticles on Proliferation and Apoptosis in Granulosa Cells of Chicken Preovulatory Follicles: An In Vitro Study

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1652
Author(s):  
Dorota Katarzyńska-Banasik ◽  
Anna Kozubek ◽  
Małgorzata Grzesiak ◽  
Andrzej Sechman
Keyword(s):  
Silver Nanoparticles ◽  
Granulosa Cells ◽  
Caspase 3 ◽  
In Vitro Study ◽  
Poultry Production ◽  
Cell Death Pathways ◽  
Preovulatory Follicles ◽  
Proliferation And Apoptosis ◽  
Apoptosis Process

The continuous development of poultry production related to the growing demand for eggs and chicken meat makes it necessary to use modern technologies. An answer to this demand may be the use of nanotechnology in poultry farming. One of the promising nanomaterials in this field are silver nanoparticles (AgNPs), which are used as disinfectants, reducing microbial pollution and the amounts of greenhouse gases released. This study aimed to evaluate the effect of AgNPs on the proliferation and apoptosis process in the granulosa cells of chicken preovulatory follicles. The in vitro culture experiment revealed that both 13 nm and 50 nm AgNPs inhibited the proliferation of the granulosa cells. However, a faster action was observed in 50 nm AgNPs than in 13 nm ones. A size-dependent effect of AgNP was also demonstrated for the caspase-3 activity. AgNPs 13 nm in size increased the caspase-3 activity in granulosa cells, while 50 nm AgNPs did not exert an effect, which may indicate the induction of distinct cell death pathways by AgNPs. In conclusion, our study reveals that AgNPs in vitro inhibit granulosa cell proliferation and stimulate their apoptosis. These results suggest that AgNPs may disrupt the final stage of preovulatory follicle maturation and ovulation.

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